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RabMAb

Anti-Cdk4抗体[EPR4513-54-3] (ab108355)

概述

  • 产品名称
    Anti-Cdk4抗体[EPR4513-54-3]
    参阅全部 Cdk4 一抗
  • 描述
    兔单克隆抗体[EPR4513-54-3] to Cdk4
  • 经测试应用
    适用于: WB, ICC/IFmore details
    不适用于: IHC-P
  • 种属反应性
    与反应: Human
  • 免疫原

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human Cdk4 aa 250 to the C-terminus (C terminal).
    Database link: P11802

  • 阳性对照
    • WB: HeLa, MCF7 and K562 cell lysates. ICC/IF: Wild-type HAP1 and HeLa cells.
  • 常规说明

    Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant rabbit monoclonal antibody.

性能

应用

Our Abpromise guarantee covers the use of ab108355 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

应用 Ab评论 说明
WB 1/1000 - 1/10000. Detects a band of approximately 34 kDa (predicted molecular weight: 34 kDa).
ICC/IF 1/400.

This product gave a positive signal in HAP1 cells fixed with 4% formaldehyde (10 min) and 100% methanol (5 min)

  • 应用说明
    Is unsuitable for IHC-P.
  • 靶标

    • 功能
      Ser/Thr-kinase component of cyclin D-CDK4 (DC) complexes that phosphorylate and inhibit members of the retinoblastoma (RB) protein family including RB1 and regulate the cell-cycle during G(1)/S transition. Phosphorylation of RB1 allows dissociation of the transcription factor E2F from the RB/E2F complexes and the subsequent transcription of E2F target genes which are responsible for the progression through the G(1) phase. Hypophosphorylates RB1 in early G(1) phase. Cyclin D-CDK4 complexes are major integrators of various mitogenenic and antimitogenic signals. Also phosphorylates SMAD3 in a cell-cycle-dependent manner and represses its transcriptional activity. Component of the ternary complex, cyclin D/CDK4/CDKN1B, required for nuclear translocation and activity of the cyclin D-CDK4 complex.
    • 疾病相关
      Defects in CDK4 are a cause of susceptibility to cutaneous malignant melanoma type 3 (CMM3) [MIM:609048]. Malignant melanoma is a malignant neoplasm of melanocytes, arising de novo or from a pre-existing benign nevus, which occurs most often in the skin but also may involve other sites.
    • 序列相似性
      Belongs to the protein kinase superfamily. CMGC Ser/Thr protein kinase family. CDC2/CDKX subfamily.
      Contains 1 protein kinase domain.
    • 翻译后修饰
      Phosphorylation at Thr-172 is required for enzymatic activity. Phosphorylated, in vitro, at this site by CCNH-CDK7, but, in vivo, appears to be phosphorylated by a proline-directed kinase. In the cyclin D-CDK4-CDKN1B complex, this phosphorylation and consequent CDK4 enzyme activity, is dependent on the tyrosine phosphorylation state of CDKN1B. Thus, in proliferating cells, CDK4 within the complex is phosphorylated on Thr-172 in the T-loop. In resting cells, phosphorylation on Thr-172 is prevented by the non-tyrosine-phosphorylated form of CDKN1B.
    • 细胞定位
      Cytoplasm. Nucleus. Membrane. Cytoplasmic when non-complexed. Forms a cyclin D-CDK4 complex in the cytoplasm as cells progress through G(1) phase. The complex accumulates on the nuclear membrane and enters the nucleus on transition from G(1) to S phase. Also present in nucleoli and heterochromatin lumps. Colocalizes with RB1 after release into the nucleus.
    • Information by UniProt
    • 数据库链接
    • 别名
      • Cdk 4 antibody
      • cdk4 antibody
      • CDK4 protein antibody
      • CDK4_HUMAN antibody
      • Cell division kinase 4 antibody
      • Cell division protein kinase 4 antibody
      • CMM 3 antibody
      • CMM3 antibody
      • Crk3 antibody
      • Cyclin dependent kinase 4 antibody
      • Cyclin-dependent kinase 4 antibody
      • Melanoma cutaneous malignant 3 antibody
      • MGC14458 antibody
      • p34 cdk4 antibody
      • PSK J3 antibody
      • PSK-J3 antibody
      see all

    Anti-Cdk4 antibody [EPR4513-54-3] 图像



    • Predicted band size : 34 kDa

      Lanes 1, 3 and 5: Wild-type HAP1 cell lysate (20 µg)
      Lanes 2, 4 and 6: CDK4 knockout HAP1 cell lysate (20 µg)
      Lanes 1 and 2: Green signal from target - ab108355 observed at 34 kDa
      Lanes 3 and 4: Red signal from loading control - ab8226 observed at 42 kDa
      Lanes 5 and 6: Merged (red and green) signal
      ab108355 was shown to specifically react with CDK4 when CDK4 knockout samples were used. Wild-type and CDK4 knockout samples were subjected to SDS-PAGE. ab108355 and ab8226 (loading control to beta actin) were both diluted at 1/1000 and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10 000 dilution for 1 h at room temperature before imaging.

    • ab108355 staining Cdk4 in wild-type HAP1 cells (top panel) and Cdk4 knockout HAP1 cells (bottom panel). The cells were fixed with 100% methanol (5min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab108355 at 1/400 dilution and ab7291 at 1μg/ml concentration overnight at +4°C, followed by a further incubation at room temperature for 1h with a goat secondary antibody to Rabbit IgG (Alexa Fluor® 488) (ab150081) at 2 μg/ml (shown in green) and a goat secondary antibody to Mouse IgG (Alexa Fluor® 594) (ab150120) (shown in pseudo colour red). Nuclear DNA was labelled in blue with DAPI.

    • Immunofluorescence staining of MCF7 cells with purified ab108355 at a working dilution of 1/100, counter-stained with DAPI. The secondary antibody was Alexa Fluor® 488 goat anti-rabbit (ab150077), used at a dilution of 1/1000. ab7291, a mouse anti-tubulin antibody (1/1000), was used to stain tubulin along with ab150120 (Alexa Fluor® 594 goat anti-mouse, 1/1000), shown in the top right hand panel. The cells were fixed in 4% PFA and permeabilized using 0.1% Triton X 100. The negative controls are shown in bottom middle and right hand panels - for negative control 1, purified ab108355 was used at a dilution of 1/500 followed by an Alexa Fluor® 594 goat anti-mouse antibody (ab150120) at a dilution of 1/500. For negative control 2, ab7291 (mouse anti-tubulin) was used at a dilution of 1/500 followed by an Alexa Fluor® 488 goat anti-rabbit antibody (ab150077) at a dilution of 1/400.

    • All lanes : Anti-Cdk4 antibody [EPR4513-54-3] (ab108355) at 1/5000 dilution (purified)

      Lane 1 : HeLa cell lysate
      Lane 2 : K562 cell lysate
      Lane 3 : Ramos cell lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      HRP goat anti-rabbit IgG (H+L) at 1/50000 dilution

      Predicted band size : 34 kDa
      Observed band size : 34 kDa

      Blocking buffer: 5% NFDM/TBST
      Dilution buffer: 5% NFDM/TBST

    • All lanes : Anti-Cdk4 antibody [EPR4513-54-3] (ab108355) at 1/1000 dilution (unpurified)

      Lane 1 : Human osteosarcoma cells - control, non-targeting siRNA
      Lane 2 : Human osteosarcoma cells - siRNA for CDK4

      Lysates/proteins at 20 µg per lane.

      Secondary
      HRP-conjugated goat anti-rabbit IgG polyclonal at 1/2000 dilution
      Developed using the ECL technique

      Performed under reducing conditions.

      Predicted band size : 34 kDa
      Observed band size : 34 kDa


      Exposure time : 5 seconds

      This image is courtesy of an Abreview submitted by Sonia Rocha

      See Abreview

    Anti-Cdk4 antibody [EPR4513-54-3] (ab108355)参考文献

    ab108355 has not yet been referenced specifically in any publications.

    Product Wall

    Application
    Western blot
    Loading amount
    20 µg
    Gel Running Conditions
    Reduced Denaturing (10%)
    Sample
    Human Cell lysate - whole cell (osteosarcoma cells)
    Specification
    osteosarcoma cells
    Treatment
    lane 1 control, lane 2 siRNA for CDK4, 48h
    Blocking step
    Milk as blocking agent for 10 minute(s) · Concentration: 10% · Temperature: 21°C
    Username

    Dr. Sonia Rocha

    Verified customer

    提交于 Sep 26 2014

    Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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